Genetic distribution of 295 Bacillus cereus group members based on adk-screening in combination with MLST (Multilocus Sequence Typing) used for validating a primer targeting a chromosomal locus in B. anthracis

Olsen, Jaran Strand; Skogan, Gunnar; Fykse, Else Marie; Rawlinson, Elizabeth L.; Tomaso, Herbert; Granum, Per Einar; Blatny, Janet Martha

doi:10.1016/j.mimet.2007.10.001

Cited by 23 publications

(18 citation statements)

References 54 publications

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“…Primers were designed to amplify internal fragments of candidate-genes of the publicly available B. licheniformis ATCC14580 genome (GenBank: NC_00627) using the Primer3 software [43]. The choice of candidate-genes was based previously published genotyping schemes for members of the Bacillus genus [28,32,36]. The primers targeted 400-718 bp fragments of the nine house-keeping genes adk , ccpA , glpT, gyrB, pyrE , recF , rpoB, sucC and spo0A which were dispersed over the entire genome .…”

Section: Methodsmentioning

confidence: 99%

“…It has been applied to numerous species including members of the B. cereus family and Clostridium spp. [32-36] and has been used for epidemiological purposes identifying strains that could cause human infections [37,38]. Basically, it relies on the sequence of several (usually six to eight) conserved house-keeping genes which are independently distributed in the genome.…”

Section: Introductionmentioning

confidence: 99%

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Genotyping of B. licheniformisbased on a novel multi-locus sequence typing (MLST) scheme

et al. 2012

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BackgroundBacillus licheniformis has for many years been used in the industrial production of enzymes, antibiotics and detergents. However, as a producer of dormant heat-resistant endospores B. licheniformis might contaminate semi-preserved foods. The aim of this study was to establish a robust and novel genotyping scheme for B. licheniformis in order to reveal the evolutionary history of 53 strains of this species. Furthermore, the genotyping scheme was also investigated for its use to detect food-contaminating strains.ResultsA multi-locus sequence typing (MLST) scheme, based on the sequence of six house-keeping genes (adk, ccpA, recF, rpoB, spo0A and sucC) of 53 B. licheniformis strains from different sources was established. The result of the MLST analysis supported previous findings of two different subgroups (lineages) within this species, named “A” and “B” Statistical analysis of the MLST data indicated a higher rate of recombination within group “A”. Food isolates were widely dispersed in the MLST tree and could not be distinguished from the other strains. However, the food contaminating strain B. licheniformis NVH1032, represented by a unique sequence type (ST8), was distantly related to all other strains.ConclusionsIn this study, a novel and robust genotyping scheme for B. licheniformis was established, separating the species into two subgroups. This scheme could be used for further studies of evolution and population genetics in B. licheniformis.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Genotyping of B. licheniformisbased on a novel multi-locus sequence typing (MLST) scheme

et al. 2012

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“…30 These genetic markers provide limited specificity and require additional timeconsuming and labor-intensive post-PCR analysis steps. Other areas of the chromosome have also been investigated as potential DNA-targets for identification purposes, including the so-called BA813 [31][32][33][34][35][36][37][38] and BA5510 sequences, 19 genes bclB, 39 sap, 40,41 saspB, 5,42 and sspE, 22,43 the B-type small acid-soluble spore protein gene (SASP), 44 a glycosyltransferase group 1 family protein, 45 a protein showing similarities with an abhydrolase, 18 and several DNA loci located on prophage regions, 17 i.e., BA5345, 21 BA5357, 46 and PL3. 47 Although most of these regions have been claimed to be anthrax-specific, B. cereus strains sometimes yield false positive results.…”

Section: Literature Survey Of Pcr-based Detection Methodsmentioning

confidence: 99%

“…[13][14][15] Few chromosomal sequences that provide sufficient polymorphism to unambiguously distinguish B. anthracis from its near neighbors have been identified. 14,[16][17][18][19][20][21][22] Some of these assays rely upon single-nucleotide differences for discrimination and are therefore sensitive to assay conditions and PCR cycling parameters. Small alterations in these conditions can result in the loss of specificity, especially with hydrolysis probes, i.e., TaqMan chemistry.…”

Section: Introductionmentioning

confidence: 99%

In silico and in vitro evaluation of PCR-based assays for the detection ofBacillus anthracischromosomal signature sequences

et al. 2013

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“…In September and October 2001 letters containing B. anthracis spores were mailed to several US locations as bioterrorist attacks [18], [19]. The closely related bacterium Bacillus thuringiensis [20] is a common plant protection agent that is pathogenic for insects, but non-pathogenic for vertebrates including humans. Therefore, B. thuringiensis has frequently been suggested as substitute of B. anthracis for field exercises.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of a microfluidic chip system for preparation of bacterial DNA from swabs, air, and surface water samples

et al. 2016

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The detection of bacterial pathogens from complex sample matrices by PCR requires efficient DNA extraction. In this study, a protocol for extraction and purification of DNA from swabs, air, and water samples using a microfluidic chip system was established. The optimized protocol includes a combination of thermal, chemical and enzymatic lysis followed by chip-based DNA purification using magnetic particles. The procedure was tested using Gram-positive Bacillus thuringiensis Berliner var. kurstaki as a model organism for Bacillus anthracis and the attenuated live vaccine strain of Francisella tularensis subsp. holarctica as Gram-negative bacterium. The detection limits corresponded to 103 genome equivalents per milliliter (GE/ml) for surface water samples spiked with F. tularensis and 102 GE/ml for B. thuringiensis. In air, 10 GE of F. tularensis per 10 L and 1 GE of B. thuringiensis per 10 L were detectable. For swab samples obtained from artificially contaminated surfaces the detection limits were 4 × 103 GE/cm2 for F. tularensis and 4 × 102 GE/cm2 for B. thuringiensis. Suitability of the chip-assisted procedure for DNA preparation of real samples was demonstrated using livestock samples. The presence of thermophilic Campylobacter spp. DNA could be confirmed in air samples collected on pig and broiler farms.

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Genetic distribution of 295 Bacillus cereus group members based on adk-screening in combination with MLST (Multilocus Sequence Typing) used for validating a primer targeting a chromosomal locus in B. anthracis

Cited by 23 publications

References 54 publications

Genotyping of B. licheniformisbased on a novel multi-locus sequence typing (MLST) scheme

Genotyping of B. licheniformisbased on a novel multi-locus sequence typing (MLST) scheme

In silico and in vitro evaluation of PCR-based assays for the detection ofBacillus anthracischromosomal signature sequences

Evaluation of a microfluidic chip system for preparation of bacterial DNA from swabs, air, and surface water samples

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